| 引用本文: |
刘卓琳,冯芷莹,王康宇,黄家望,时佳宁,张成,李玲,何清湖.基于HIF-1α/EGFR/MAPK1信号通路探讨龟鹿二仙胶对少弱精子症大鼠的保护作用[J].湖南中医药大学学报,2024,44(4):572-579[点击复制] |
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| 基于HIF-1α/EGFR/MAPK1信号通路探讨龟鹿二仙胶对少弱精子症大鼠的保护作用 |
| 刘卓琳,冯芷莹,王康宇,黄家望,时佳宁,张成,李玲,何清湖 |
| (湖南中医药大学中西医结合学院, 湖南 长沙 410208;湖南中医药大学中医学院, 湖南 长沙 410208;湖南中医药大学男科实验室, 湖南 长沙 410208;湖南医药学院中医学院, 湖南 怀化 418000) |
| 摘要: |
| 目的 探讨龟鹿二仙胶调控缺氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）/表皮生长因子受体（epidermal growth factor receptor，EGFR）/和丝裂原活化蛋白激酶（mitogen-activated protein kinase 1，MAPK1）信号通路对少弱精子症（oligoasthenozoospermia，OAS）大鼠的保护作用。方法 将48只SD雄性大鼠随机分为正常组，模型对照组，龟鹿二仙胶低剂量（0.65 g/kg）组、中剂量（1.25 g/kg）组、高剂量（2.5 g/kg）组和左卡尼汀组，每组8只。除正常组外，其他组均采用腺嘌呤灌胃给药进行造模。造模成功后进行灌胃给药进行干预，连续给药4周，记录大鼠一般情况，给药4周后进行取材处理。腹主动脉采血，ELISA法检测血清睾酮（testosterone，T）、雌二醇（estradiol，E2）、促卵泡激素（follicle-stimulating hormone，FSH）、促黄体生成激素（luteinizing hormone，LH）和催乳素（prolactin，PRL）；分离肾组织，计算肾指数；分离右侧附睾，运用自动精子分析仪检测各组大鼠精子浓度和精子活力；HE染色观察睾丸组织病理改变；用试剂盒检测大鼠睾丸组织活性氧（reactive oxygen species，ROS）、超氧化物歧化酶（superoxide dismutase，SOD）和谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）水平；免疫组化检测大鼠睾丸组织HIF-1α、EGFR和MAPK1的蛋白表达。结果 与模型对照组相比，不同剂量的龟鹿二仙胶均可显著增加大鼠的体质量（P＜0.01），降低大鼠的肾指数（P＜0.01），提高精子浓度和精子活力（P＜0.01），改善大鼠的精子质量，并能有效减轻模型大鼠睾丸组织损伤程度，其中与龟鹿二仙胶低剂量组相比，龟鹿二仙胶高剂量组大鼠体质量、精子密度和精子活力均显著上升（P＜0.05）。ELISA结果显示，与模型对照组相比，不同剂量龟鹿二仙胶能显著升高T和E2水平（P＜0.05），且显著降低FSH、LH和PRL水平（P＜0.05）。ROS、SOD和GSH-Px检测结果显示，与模型对照组相比，龟鹿二仙胶低、中、高剂量组ROS水平显著降低（P＜0.05），龟鹿二仙胶中、高剂量组和左卡尼汀组SOD和GSH-Px活性显著升高（P＜0.05）。免疫组化结果显示，与模型对照组相比，龟鹿二仙胶中、高剂量组和左卡尼汀组大鼠睾丸组织HIF-1α、EGFR、MAPK1水平显著降低（P＜0.05）。结论 龟鹿二仙胶可能是通过HIF-1α/EGFR/MAPK1信号通路调节氧化应激来改善OAS。 |
| 关键词: 少弱精子症 龟鹿二仙胶 HIF-1α/EGFR/MAPK1信号通路 精子活力 氧化应激 |
| DOI：10.3969/j.issn.1674－070X.2024.04.007 |
| 投稿时间：2023-09-12 |
| 基金项目:国家自然科学基金项目（81973863）；湖南省大学生创新创业训练计划项目（2022-2864）。 |
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| Protective effects of Guilu Erxian Glue on rats with oligoasthenospermia based on HIF-1α/EGFR/MAPK1 signaling pathway |
| LIU Zhuolin,FENG Zhiying,WANG Kangyu,HUANG Jiawang,SHI Jianing,ZHANG Cheng,LI Ling,HE Qinghu |
| (College of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;College of Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Andrology Laboratory, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;College of Chinese Medicine, Hunan University of Medicine, Huaihua, Hunan 418000, China) |
| Abstract: |
| Objective To explore the protective effects of Guilu Erxian Glue (GLEXG) on rats with oligoasthenospermia (OAS) by regulating the hypoxia inducible factor-1α/epidermal growth factor receptor/mitogen-activated protein kinase 1(HIF-1α/EGFR/MAPK1) signaling pathway.Methods Forty-eight male SD rats were randomized into normal,model control,low-dose GLEXG (0.65 g/kg),medium-dose GLEXG (1.25 g/kg),high-dose GLEXG (2.5 g/kg),and L-carnitine groups,with eight rats in each group.All groups except the normal group were administered adenine by gavage to create the model After successful modeling,the rats were intervened by intragastric administration for continuous four weeks,during which the general condition of the rats was recorded.After four weeks of administration,the rats were processed for sample collection.Blood was collected from the abdominal aorta.Serum testosterone (T),estradiol (E2),follicle-stimulating hormone (FSH),luteinizing hormone (LH),and prolactin (PRL) levels were measured by ELISA.Renal tissues were separated to calculate the renal index.The right epididymis was separated,and the sperm concentration and motility of each group were checked by automatic sperm analyzer.The pathological changes of testicular tissue were observed by HE staining.Reactive oxygen species (ROS),superoxide dismutase (SOD),and glutathione peroxidase (GSH-Px) levels in rat testicular tissue were examined by respective assay kits.Protein expressions of HIF-1α,EGFR,and MAPK1 in rat testicular tissue were checked by immunohistochemistry.Results Compared with the model control group,different doses of GLEXG significantly increased the body mass of rats (P＜0.01),decreased their renal index of rats (P＜0.01),improved sperm concentration and motility (P＜0.01),thereby enhancing the sperm quality of rats and effectively reducing the degree of testicular tissue damage in the model rats.Compared with the low-dose GLEXG group,the high-dose group showed significant increase in body mass,sperm density,and sperm motility (P＜0.05).ELISA results showed that compared with model control group,different doses of GLEXG significantly increased T and E2 levels (P＜0.05),and significantly decreased FSH,LH,and PRL levels (P＜0.05).The results of ROS,SOD,and GSH-Px indicated that compared with the model control group,ROS levels in low-,medium-,and high-dose GLXEG groups were significantly lower (P＜0.05),while SOD and GSH-Px activities in medium-and high-dose GLXEG,and L-carnitine groups were significantly higher (P＜0.05).Immunohistochemical results showed that compared with the model control group,the levels of HIF-1α,EGFR,and MAPK1 in rat testicular tissue of medium-,high-dose GLXEG and L-carnitine groups were significantly lower (P＜0.05).Conclusion GLEXG may improve OAS by regulating oxidative stress through HIF-1α/EGFR/MAPK1 signaling pathway. |
| Key words: oligoasthenospermia Guilu Erxian Glue HIF-1α/EGFR/MAPK1 signaling pathway sperm motility oxidative stress |
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